Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1973 Sep;12(3):600–608. doi: 10.1128/jvi.12.3.600-608.1973

Hydroxyurea-Induced Accumulation of Short Fragments During Polyoma DNA Replication I. Characterization of Fragments

Göran Magnusson 1
PMCID: PMC356668  PMID: 4355857

Abstract

Hydroxyurea treatment of 3T6 mouse fibroblast cells infected with polyoma virus resulted within 15 min in more than a 20-fold reduction of the rate of both viral and cellular DNA synthesis. After the initial rapid inhibition, the rate of DNA synthesis remained essentially constant for at least 2 h. In the inhibited cells viral DNA accumulated as short chains with a sedimentation coefficient of about 4S (hydroxyurea fragments). A variable proportion of these fragments was released from the template strands when the viral DNA was extracted by the Hirt procedure. Reannealing experiments demonstrated that hydroxyurea fragments were polyoma-specific and probably synthesized on both parental strands at the replication forks.

Full text

PDF
602

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bourgaux P., Bourgaux-Ramoisy D., Dulbecco R. The replication of the ring-shaped DNA of polyoma virus. I. Identification of the replicative intermediate. Proc Natl Acad Sci U S A. 1969 Oct;64(2):701–708. doi: 10.1073/pnas.64.2.701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Eckhart W. Complementation and transformation by temperature-sensitive mutants of polyoma virus. Virology. 1969 May;38(1):120–125. doi: 10.1016/0042-6822(69)90133-0. [DOI] [PubMed] [Google Scholar]
  3. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  4. Krakoff I. H., Brown N. C., Reichard P. Inhibition of ribonucleoside diphosphate reductase by hydroxyurea. Cancer Res. 1968 Aug;28(8):1559–1565. [PubMed] [Google Scholar]
  5. LEHMAN I. R., NUSSBAUM A. L. THE DEOXYRIBONUCLEASES OF ESCHERICHIA COLI. V. ON THE SPECIFICITY OF EXONUCLEASE I (PHOSPHODIESTERASE). J Biol Chem. 1964 Aug;239:2628–2636. [PubMed] [Google Scholar]
  6. Levine A. J., Kang H. S., Billheimer F. E. DNA replication in SV40 infected cells. I. Analysis of replicating SV40 DNA. J Mol Biol. 1970 Jun 14;50(2):549–568. doi: 10.1016/0022-2836(70)90211-1. [DOI] [PubMed] [Google Scholar]
  7. Magnusson G., Pigiet V., Winnacker E. L., Abrams R., Reichard P. RNA-linked short DNA fragments during polyoma replication. Proc Natl Acad Sci U S A. 1973 Feb;70(2):412–415. doi: 10.1073/pnas.70.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Magnusson G., Winnacker E. L., Eliasson R., Reichard P. Replication of polyoma DNA in isolated nuclei. II. Evidence for semi-conservative replication. J Mol Biol. 1972 Dec 30;72(3):539–552. doi: 10.1016/0022-2836(72)90173-8. [DOI] [PubMed] [Google Scholar]
  9. Meinke W., Goldstein D. A. Studies on the structure and formation of polyoma DNA replicative intermediates. J Mol Biol. 1971 Nov 14;61(3):543–563. doi: 10.1016/0022-2836(71)90064-7. [DOI] [PubMed] [Google Scholar]
  10. Moore E. C. The effects of ferrous ion and dithioerythritol on inhibition by hydroxyruea of ribonucleotide reductase. Cancer Res. 1969 Feb;29(2):291–295. [PubMed] [Google Scholar]
  11. Oishi M. Studies of DNA replication in vivo. I. Isolation of the first intermediate of DNA replication in bacteria as single-stranded DNA. Proc Natl Acad Sci U S A. 1968 May;60(1):329–336. doi: 10.1073/pnas.60.1.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Okazaki R., Okazaki T., Sakabe K., Sugimoto K., Sugino A. Mechanism of DNA chain growth. I. Possible discontinuity and unusual secondary structure of newly synthesized chains. Proc Natl Acad Sci U S A. 1968 Feb;59(2):598–605. doi: 10.1073/pnas.59.2.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. RICHARDSON C. C., KORNBERG A. A DEOXYRIBONUCLEIC ACID PHOSPHATASE-EXONUCLEASE FROM ESCHERICHIA COLI. I. PURIFICATION OF THE ENZYME AND CHARACTERIZATION OF THE PHOSPHATASE ACTIVITY. J Biol Chem. 1964 Jan;239:242–250. [PubMed] [Google Scholar]
  14. RICHARDSON C. C., LEHMAN I. R., KORNBERG A. A DEOXYRIBONUCLEIC ACID PHOSPHATASE-EXONUCLEASE FROM ESCHERICHIA COLI. II. CHARACTERIZATION OF THE EXONUCLEASE ACTIVITY. J Biol Chem. 1964 Jan;239:251–258. [PubMed] [Google Scholar]
  15. Sebring E. D., Kelly T. J., Jr, Thoren M. M., Salzman N. P. Structure of replicating simian virus 40 deoxyribonucleic acid molecules. J Virol. 1971 Oct;8(4):478–490. doi: 10.1128/jvi.8.4.478-490.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Skoog L., Nordenskjöld B. Effects of hydroxyurea and 1-beta-D-arabinofuranosyl-cytosine on deoxyribonucleotide pools in mouse embryo cells. Eur J Biochem. 1971 Mar 1;19(1):81–89. doi: 10.1111/j.1432-1033.1971.tb01290.x. [DOI] [PubMed] [Google Scholar]
  17. Sugino A., Hirose S., Okazaki R. RNA-linked nascent DNA fragments in Escherichia coli. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1863–1867. doi: 10.1073/pnas.69.7.1863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Thoren M. M., Sebring E. D., Salzman N. P. Specific initiation site for simian virus 40 deoxyribonucleic acid replication. J Virol. 1972 Sep;10(3):462–468. doi: 10.1128/jvi.10.3.462-468.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Vinograd J., Lebowitz J., Radloff R., Watson R., Laipis P. The twisted circular form of polyoma viral DNA. Proc Natl Acad Sci U S A. 1965 May;53(5):1104–1111. doi: 10.1073/pnas.53.5.1104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Winnacker E. L., Magnusson G., Reichard P. Replication of polyoma DNA in isolated nuclei. I. Characterization of the system from mouse fibroblast 3T6 cells. J Mol Biol. 1972 Dec 30;72(3):523–537. doi: 10.1016/0022-2836(72)90172-6. [DOI] [PubMed] [Google Scholar]
  21. YOUNG C. W., HODAS S. HYDROXYUREA: INHIBITORY EFFECT ON DNA METABOLISM. Science. 1964 Nov 27;146(3648):1172–1174. doi: 10.1126/science.146.3648.1172. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES